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 Table of Contents  
ORIGINAL ARTICLE
Year : 2023  |  Volume : 20  |  Issue : 1  |  Page : 20

Evaluation of the antimicrobial efficacy of different concentrations of a novel root canal filling material for primary teeth - An in vitro study


Department of Paediatric and Preventive Dentistry, Saveetha Dental College and Hospitals, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu, India

Date of Submission27-May-2022
Date of Acceptance15-Dec-2022
Date of Web Publication14-Feb-2023

Correspondence Address:
Dr. Lavanya Govindaraju
Department of Paediatric and Preventive Dentistry, Saveetha Dental College and Hospitals, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/1735-3327.369622

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  Abstract 


Background: The demand for an ideal root canal filling material in primary teeth with maximum antibacterial properties keeps escalating to improve the success rate of root canal treatment in primary teeth. Hence, the present study was conducted to determine the antimicrobial efficacy of the different concentrations of a novel root canal filling material.
Materials and Methods: An in vitro study was designed to compare the antimicrobial efficacy of different concentrations of the novel obturating material for primary teeth. Different concentrations of the novel obturating material were prepared using calcium hydroxide, zinc oxide cement, and metronidazole and were tested for its antimicrobial efficacy against Streptococcus mutans and Enterococcus faecalis using Mueller–Hinton Agar for 24 h at 37°C. The zone of inhibition was measured in millimeters after 24 h. The values were entered into a Microsoft Excel spreadsheet and subjected to statistical analysis. Descriptive statistics were used for data summarization and presentation. Friedman test and Kruskal–Wallis test were used to compare the different concentrations of the novel obturating materials against S. mutans and E. faecalis. P < 0.05 was considered to be significant.
Results: Against S. mutans at 1% concentration, the zone of inhibition was statistically greater at 50 μl and at 2% and 3%, the zone of inhibition was greater at 100 μl. Similarly, against E. faecalis, the zone of inhibition was statistically greater at 100 μl at 2% and no significant difference was noted at 1% and 3%. Comparison between the groups at 25, 50, and 100 μl shows no significant difference against S. mutans between 1%, 2%, and 3% of 60–40 concentration. However, 3% 60–40 concentration at 50 μl shows significantly increased zone of inhibition against E. faecalis.
Conclusion: Two percentage metronidazole at 60–40 (calcium hydroxide–zinc oxide) concentration of the novel obturating material at 100 μl has better antibacterial properties against both S. mutans and E. faecalis.

Keywords: Child, deciduous, in vitro techniques, microbiology, pulpectomy, tooth


How to cite this article:
Govindaraju L, Jeevanandan G. Evaluation of the antimicrobial efficacy of different concentrations of a novel root canal filling material for primary teeth - An in vitro study. Dent Res J 2023;20:20

How to cite this URL:
Govindaraju L, Jeevanandan G. Evaluation of the antimicrobial efficacy of different concentrations of a novel root canal filling material for primary teeth - An in vitro study. Dent Res J [serial online] 2023 [cited 2023 Sep 23];20:20. Available from: https://www.drjjournal.net/text.asp?2023/20/1/20/369622




  Introduction Top


Root canal treatment is the only option to treat primary teeth with pulpal signs and symptoms.[1] However, root canal treatment in primary teeth is a strenuous procedure owing to the complex and torturous nature of the root canal system. Despite proper canal preparation and irrigation in the primary teeth, the microorganisms still remain entrapped leading to failure of the root canal treatment.[2],[3] Hence, the success of root canal treatment in primary teeth majorly depends on the obturating material used. An ideal root canal filling material in primary teeth should have maximal antibacterial activity and at the same time should resorb at the same rate of the primary teeth without exerting any deleterious effects on the permanent teeth and the periapical tissues.[4],[5]

Conventionally used root canal filling materials such as zinc oxide eugenol cement, calcium hydroxide, and iodoform paste has its respective disadvantages. The former material has a low resorption rate which in turn results in the deflection of the permanent teeth during its eruption.[6] While the latter material resorbs much faster than the primary teeth resulting in a hollow tube effect of the primary root canals serving as a nidus for reinfection.[6],[7] The above evidence authenticates the need for an ideal root canal filling material to proclaim the success of endodontic treatment in primary teeth.

A novel root canal filling material was developed with zinc oxide, calcium hydroxide, and metronidazole at different concentrations and was checked for antimicrobial efficacy to determine the most efficacious composition. The different concentrations of the novel root canal filling material were checked against Streptococcus mutans and Enterococcus faecalis as these organisms have been reported to be present in higher concentrations in necrotic primary teeth.[8],[9] The present study aimed to determine the antimicrobial efficacy of the different concentrations of the novel root canal filling material against S. mutans and E. faecalis in primary teeth and also to determine the most efficacious composition of the novel root canal filling material.


  Materials and Methods Top


An in vitro study was designed to compare the antimicrobial efficacy of different concentrations of the novel obturating material for primary teeth.

Preparation of the obturating material

Six hundred milligrams of calcium hydroxide powder was mixed with 400 mg of zinc oxide powder to form 60–40 concentration of the mixture and was kept in a magnetic stirrer for 1 h. Four hundred milligrams of metronidazole tablet was crushed into powder using motor and pistle. One percentage of calcium hydroxide–zinc oxide–metronidazole powder was prepared by mixing 2 mg of metronidazole powder to 198 mg of the calcium hydroxide–zinc oxide mixture. Similarly, 2% and 3% calcium hydroxide–zinc oxide–metronidazole powder were prepared by mixing 4 and 6 mg of metronidazole powder to 196 and 194 mg of zinc oxide–calcium hydroxide mixture, respectively. The samples were mixed with 2 μl of distilled water, placed in the vortex for 15 min, and then in a mixer for 1 day.

Preparation of the culture medium

The antimicrobial activity of the different concentrations of the novel obturating material against S. mutans and E. faecalis was determined using Mueller–Hinton Agar (MHA). MHA was prepared and sterilized at 121° C for 15 min. The media was poured into 30 sterilized plates (15 for S. mutans and 15 for E. faecalis) and was allowed for solidification. The wells were cut using the well cutter, and the test organisms were swabbed. 1%, 2%, and 3% of the novel obturating material were loaded at 25, 50, and 100 μl and were incubated at 37°C for 1 day. The zone of inhibition was measured in millimeters after 24 h of incubation by measuring the shortest distance between the outer margin of the well and initial microbial growth.

Statistical analysis

Data were entered in a Microsoft Excel spreadsheet and analyzed using SPSS software (IBM SPSS Statistics, Version 20.0, Arming, NY, USA: IBM Corp.). Descriptive statistics were used for data summarization and presentation. Friedman test and Kruskal–Wallis test were used to compare the different concentrations of the novel obturating materials against S. mutans and E. faecalis.


  Results Top


In the present study with 60–40 concentrations of the novel obturating material, it was seen that at 1%, the zone of inhibition was statistically greater at 50 μl against S. mutans. However, at 2% and 3%, the zone of inhibition was greater at 100 μl against S. mutans, the former being statistically significant. Similarly, against E. faecalis, the zone of inhibition was statistically greater at 100 μl at 2% and no significant difference was noted at 1% and 3% [Table 1].
Table 1: Comparison of different concentrations of the novel obturating material against Streptococcus mutans and Enterococcus faecalis

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Comparison between the groups at 25, 50, and 100 μl shows no significant difference against S. mutans between 1%, 2%, and 3% of 60–40 concentration. However, 3% 60–40 concentration at 50 μl shows a significantly increased zone of inhibition against E. faecalis [Table 2].
Table 2: Comparison between 1%, 2%, and 3% 60-40 concentration of the novel obturating material against Streptococcus mutans and Enterococcus faecalis at 25, 50, and 100 μl

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  Discussion Top


A systematic review comparing zinc oxide eugenol and calcium hydroxide/iodoform paste as root canal filling material for pulpectomy in primary teeth concluded that the combination of zinc oxide eugenol and calcium hydroxide/iodoform paste is the material of choice for root canal treatment in primary teeth.[10] Despite the fact that the antimicrobial activity of zinc oxide eugenol is mainly attributed to eugenol, it can cause irritation to the periapical region, necrosis of the bone, and cementum.[11],[12] Similarly, with iodoform, yellowish discoloration and accelerated resorption of the primary teeth were reported.[5],[13] Hence, a novel root canal filling material was developed by substituting eugenol and iodoform, which are known for its antibacterial properties with metronidazole to the calcium hydroxide and zinc oxide powder (60–40 concentration). 1%, 2%, and 3% metronidazole were added to 60–40 calcium hydroxide and zinc oxide mixture and were evaluated for its antimicrobial efficacy against S. mutans and E. faecalis to certify the most efficacious concentration of the novel root canal filling material.

The results of the present study show that against S. mutans, at 1% concentration, the zone of inhibition is statistically greater at 50 μl. However, the zone of inhibition was greater for E. faecalis only at 100 μl with 1% concentration of the novel root canal filling material. At 2% 60–40 concentration, the zone of inhibition was statistically greater at 100 μl for both S. mutans and E. faecalis. No significant difference was noted with 3% 60–40 concentration of the novel root canal filling material. These results authenticate that the novel root canal filling material shows better antimicrobial properties at 100 μl. Comparison in between the groups at 100 μl shows no statistically significant difference, however, at 2% concentration shows a greater zone of inhibition.

In the present study, during the development of the material, the concentration of calcium hydroxide was increased when compared to zinc oxide as calcium hydroxide itself exhibits antibacterial properties by ionic dissociation of calcium and hydroxyl ions while the antibacterial effect of zinc oxide-based materials was majorly due to eugenol. Metronidazole was selected as a substitute to eugenol and iodoform as it is known that the root canal space is majorly invaded by anaerobic bacteria and metronidazole fights against anaerobic facultative microorganisms by diffusing into the organism and inhibiting the protein synthesis causing cell death of the susceptible organisms.[14]


  Conclusion Top


From the results of the present study, it can be concluded that 2% metronidazole at 60–40 (calcium hydroxide–zinc oxide) concentration of the novel obturating material at 100 μl has better antibacterial properties against both S. mutans and E. faecalis.

Financial support and sponsorship

Nil.

Conflicts of interest

The authors of this manuscript declare that they have no conflicts of interest, real or perceived, financial or non-financial in this article.



 
  References Top

1.
Kubota K, Golden BE, Penugonda B. Root canal filling materials for primary teeth: A review of the literature. ASDC J Dent Child 1992;59:225-7.  Back to cited text no. 1
    
2.
American Academy of Pediatric Dentistry Clinical Affairs Committee–Pulp Therapy Subcommittee, American Academy of Pediatric Dentistry Council on Clinical Affairs. Guideline on pulp therapy for primary and young permanent teeth. Pediatr Dent 2005;27:130-4.  Back to cited text no. 2
    
3.
Bodrumlu E, Semiz M. Antibacterial activity of a new endodontic sealer against Enterococcus faecalis. J Can Dent Assoc 2006;72:637.  Back to cited text no. 3
    
4.
Rifkin A. A simple, effective, safe technique for the root canal treatment of abscessed primary teeth. ASDC J Dent Child 1980;47:435-41.  Back to cited text no. 4
    
5.
Garcia-Godoy F. Evaluation of an iodoform paste in root canal therapy for infected primary teeth. ASDC J Dent Child 1987;54:30-4.  Back to cited text no. 5
    
6.
Coll JA, Sadrian R. Predicting pulpectomy success and its relationship to exfoliation and succedaneous dentition. Pediatr Dent 1996;18:57-63.  Back to cited text no. 6
    
7.
Nurko C, Ranly DM, García-Godoy F, Lakshmyya KN. Resorption of a calcium hydroxide/iodoform paste (Vitapex) in root canal therapy for primary teeth: A case report. Pediatr Dent 2000;22:517-20.  Back to cited text no. 7
    
8.
Rifkin A. The root canal treatment of abscessed primary teeth – A three to four year follow-up. ASDC J Dent Child 1982;49:428-31.  Back to cited text no. 8
    
9.
Hegde AM, Lakshmi P. Prevalence of selected microorganisms in the pulp space of human deciduous teeth with irreversible pulpitis. Endodontology 2013;25:107-11.  Back to cited text no. 9
  [Full text]  
10.
Najjar RS, Alamoudi NM, El-Housseiny AA, Al Tuwirqi AA, Sabbagh HJ. A comparison of calcium hydroxide/iodoform paste and zinc oxide eugenol as root filling materials for pulpectomy in primary teeth: A systematic review and meta-analysis. Clin Exp Dent Res 2019;5:294-310.  Back to cited text no. 10
    
11.
Spedding RH. Incomplete resorption of resorbable zinc oxide root canal fillings in primary teeth: Report of two cases. ASDC J Dent Child 1985;52:214-6.  Back to cited text no. 11
    
12.
Hendry JA, Jeansonne BG, Dummett CO Jr., Burrell W. Comparison of calcium hydroxide and zinc oxide and eugenol pulpectomies in primary teeth of dogs. Oral Surg Oral Med Oral Pathol 1982;54:445-51.  Back to cited text no. 12
    
13.
Moskovitz M, Tickotsky N, Ashkar H, Holan G. Degree of root resorption after root canal treatment with iodoform-containing filling material in primary molars. Quintessence Int 2012;43:361-8.  Back to cited text no. 13
    
14.
Weir CB, Le JK. Metronidazole. In: StatPearls. Treasure Island (FL): StatPearls Publishing; 2022. Available from: https://www.ncbi.nlm.nih.gov/books/NBK539728/. [Last updated on 2021 Jun 29].  Back to cited text no. 14
    



 
 
    Tables

  [Table 1], [Table 2]



 

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